Therapeutic Composition of Prangos ferulacea (L.) Lindl. Essential Oil for Use in the Prevention and / or Treatment of Sepsis-Induced Acute Lung Injury

Abstract

This invention was developed to reduce the excessive inflammatory response and mitigate tissue damage observed in acute lung injury resulting from sepsis. Within the scope of this invention, it has been shown that the application of essential oil obtained from the seeds of Prangos ferulacea (L.) Lindl. in appropriate doses reduces the expression of pro-inflammatory mediators such as NF-κB, TNF-κB, IL-1β, IL-6, and HMGB1, and alleviates cellular infiltration and structural damage in alveolar tissue. This technical solution offers an alternative approach to treating sepsis-induced lung damage through the use of a naturally derived, biocompatible, and low-toxicity compound. Thus, the invention enables the development of an innovative and effective anti-inflammatory treatment strategy in inflammatory processes where current pharmacological treatments are insufficient.

Description

1 TARIFF In the Prevention and / or Treatment of Sepsis-Induced Acute Lung Injury Therapeutic Properties of Prangos ferulacea (L.) L ndl. Essential Oil for Use B leş m TECHNICAL AREA 5 The invention relates to acute pulmonary injury (ALI) and acute respiratory distress caused by sepsis. Prangos is used for the prevention and / or treatment of acute nervous system disorders (ARDS). The essential oil compounds obtained from the plant ferulacea (L.) L ndl. are pharmaceutical. and the field of biotechnology. Thus, naturally derived therapeutic compounds development and treatment of inflammatory lung diseases 10 It is also included in its applicability. STATE OF THE ART Sepsis is a common disease worldwide, and it is frequently seen in intensive care units. With very high mortality and morbidity rates, it is one of the most important public health problems. brdr (Iwashyna et al., 2010; Zeng et al., 2015). Sepsis is the most common disease affecting... Tissue of the lung. In the pathophysiology of ARDS and / or ALI resulting from sepsis. Excessive release of pro-inflammatory cytokines leads to a large number of neutrophils in the lungs. br kmes occurs with the increased production of reactive oxygen species (ROS). (Karabulut Uzunçakmak et al., 2023; Matthay et al., 2012). Sep sn pathogenesis, 20 Tumors are formed from certain immune cells, including monocytes / macrophages. Necrosis Factor-alpha (TNF-alpha), Interleukin-6 (IL-6) and IL-1-beta are pro-inflammatory agents. By releasing and secreting substances, the immune response against bacteria It is stated that development can be improved (Karamese et al., 2016). In addition, Nuclear factor kappa- Synthesis of many molecules such as NF-κB, IL-1β, IL-6 and TNF-α and other inflammatory processes 25 It regulates response-related functions, initiating and propagating the inflammatory response. It plays an important role in determining the pathophysiology of sepsis (Güler et al., 2024). Despite significant progress, treatment still involves antibiotics and aggressive liquids. Resuscitation, vasopressor administration, and supportive care are preferred. (Dell nger et al., 2013; S nger et al., 2016). 30 Sepsis, chromatoanatomical changes, lungs, the most basic functional unit of the lungs The deterioration of the alveolar-capillary barrier is the cause of the disease. Inflammatory cytokines (TNF-α, Vascular endothelial damage caused by IL-1α release, endothelial cells 2 It is formed by the breakdown of the t ght junctions between them. Therefore, in terms of protein... Edema forms due to the infiltration of rich fluid into the interstitial space (Su et al., 2024). Type I and Damage to type II pneumostats completely destroys the bar function. Type I pneumocystis damage causes fibrosis (Kasper et al., 1997), while Type II Damage to pneumoderms leads to disturbances in surface tension (Bardales et al., 1996). 5 Apaceae is a family that includes medicinal and aromatic plants, and many species are used in medicine. It is used as a raw material in industry (Sp nozz et al., 2021). Ap aceae Prangos ferulacea (L.) L ndl, belonging to the family lyasına, is mainly found in the Eastern Mediterranean. It is a species that grows in the region and in Western Asia (Badalament et al., 2022). P. ferulacea (L.) L ndl. 10 in the treatment of kidney and urinary tract diseases with the name “Ippomarathon” It has been recorded (Touwa et al., 2005). Gastrointestinal Prangos ferulacea. It is used as a softening agent, tonic, and gas in the treatment of ailments. It has sedative, antiviral and antihelminthic effects (Sadrae et al., (2012). One of the most prominent and attention-grabbing components of medicinal plants. Essential oils (EOs) are very potent, natural, and biologically active metabolites. substances (Erdoğan et al., 2023). The essential oil obtained from the roots of P. ferulacea, In addition to its strong antibacterial activity, it also stimulates the proliferation and migration of fibroblasts. Its ability to stimulate collagen production reveals promising wound healing properties. This has led to the observation that these findings support the traditional use of wound healing agents. It has been reported that its use will be supported (Yousef et al., 2017). 20 Furthermore, this essential oil increased the expression of NF-κB, TNF-α, IL-1β, IL-6, and HMGB1. by regulating and suppressing the inflammatory response and having a protective effect on lung tissue There is no technical solution currently available to demonstrate this. Sepsis is a disease resulting from the infiltration of a harmful pathogen. It is a medical condition resulting from the presence of pathological and physiological irregularities. 25 (Da et al., 2024). The most common organ damage in sepsis is ALI, and severe lung damage occurs. It causes acute lung damage (L u et al., 2022). can prevent or treat, effectively suppress the inflammatory response, There is a need for safer and more bioavailable new treatment agents. It is heard that, especially, natural compounds obtained from medicinal aromatic plants, 30 The effects of sepsis on lung damage are not yet fully understood. [Further details about the study follow.] Natural products obtained from medicinal and aromatic plants over the years have multiple biological effects. They are noteworthy because of their low prevalence and relatively low side effect profiles. 3 However, essential oils obtained from these plants are not effective against acute pulmonary disease associated with sepsis. the effects on damage, mechanisms of action and optimal dose ranges of the technique It is not included in the bl nen situation. Specifically Prangos ferulacea (L.) L ndl. Essential oil reduces the inflammatory response and lung tissue damage caused by sepsis. Scientific data on how it affects the molecular and histopathological levels are limited. 5 EXPLANATION OF THE INVENTION This invention explains the excessive inflammation seen in acute lung injury resulting from sepsis. It was developed to reduce the response and mitigate tissue damage. The invention Within this scope, volatile 10 obtained from seeds of the species Prangos ferulacea (L.) L ndl. With the application of the oil in appropriate doses, NF-κB, TNF-α, IL-1β, IL-6 and HMGB1 gb In alveolar tissue, cellular expression of pro-inflammatory mediators is reduced. It has been shown that infiltration and structural degradation are milder. This technical solution uses a naturally derived, biocompatible, and low-toxicity compound. The use of alternative fiber optic brachycardia in the treatment of pulmonary damage caused by sepsis 15 It offers a solution where current pharmacological treatments are insufficient. Innovative and effective anti-inflammatory treatment strategies in inflammatory processes. It enables its development.  Being a natural and biocompatible agent: Obtained from medicinal aromatic KBRB substances. By using edible essential oil, synthetic anti-inflammatory 20 lower toxicity and higher compatibility potential compared to other agents. presents. 1- Targeting multiple inflammatory pathways: Inflammatory pathways associated with sepsis. HMGB1, NF-κB, TNF-α, IL-1β and IL-6, which play a role in the process, are pro-inflammatory agents. mediators increase their expression simultaneously with their expression 25 by suppressing (by inhibiting) it, compared to single-target therapies. It provides comprehensive and effective anti-inflammatory effects.  It has a protective effect against lung tissue damage caused by sepsis. By reducing alveolar-capillary barrier displacement, it preserves the structural integrity of lung tissue. It reduces damage and lung damage index; thus, ALI and ARDS development is 30. It helps prevent [the spread of disease].  It has a protective effect against lung tissue damage caused by sepsis. By reducing alveolar-capillary barrier displacement, it preserves the structural integrity of lung tissue. 4 It reduces damage and lung damage index; thus improving ALI and ARDS. It helps prevent [the spread of disease].  Application variety: Pharmaceutical product, phytotherapeutic supplement or veterinary can be developed as a preparation Explanation of the Figures Fig. 1. Prangos ferulacea (L.) L ndl. Compounds of Essential Oil Figure 2. Scanning electron microscope image of lung tissue. Fig. 3. AB. Prangos ferulacea (L.) L ndl in the CLP-induced sepsis model. essential oil, lung Koh opening width and length (n:3) 10 Figure 4A. Lung histopathological appearance. Figure 4B. Lung damage. Figure 5. HMGB1, NF-kB, TNF-α, IL-1β and IL-6 in lung tissues of septic rats. expressions DETAILED DESCRIPTION OF THE INVENTION In this detailed description, the subject of the invention is sepsis-induced acute lung injury. For use in the prevention and / or treatment of prangos ferulacea (L.) l ndl. volatile The therapeutic properties of the oil are related to the subject and only contribute to a better understanding of the topic. It contains directive explanations and does not in any way contain a limiting characteristic. 20 B tk material Collection of Specimens and Fieldwork Examples of B tk taxa were collected in the Kastamonu province during 2023-2024. Collected through field studies conducted on the northern slopes of the Ilgaz Mountains, The work titled "Flora of Turkey and Flora of the Eastern Aegean Islands" (Davs, 1972) 25 This was done using [method name]. In field studies, the fruits of the plants were preferred. A total of 5000 grams of fruit samples were collected from each taxon as a result of studies lasting 40 days over 2 years. collected. Drying of Samples After the collected plant materials, especially the fruits, are brought to the laboratory, 30 from diseased parts, soil and other foreign matter under ambient conditions It was purified, cleaned, and dried for approximately five days. B tk Examples nn Granulation The dried plant materials are processed to extract essential oils immediately before water distillation. To be granulated in order to be reduced to a size that can fit into the balloon of the distillation apparatus. This process was carried out using a grinding machine (28,000 rpm), and the samples were 15. The wind has been subjected to grinding processes. Obtaining Essential Oils from Plant Samples 5 Extraction of essential oils from dried plant material, essential oil distillation equipment. The process was carried out and the resulting oils were stored at +4 °C for one week. In the water distillation method, 100 g of cleaned and ground fruit The samples are placed in a glass balloon, and the connection between the balloon and the cooler is made of silicone. The oil was used to prepare the samples. The samples were boiled at 100 °C for 3 hours, then finger-sized portions were used. The volatile oil and hydrolate phases of d-st-lat, which was condensed in the cooler, were separated. GC-MS Method (Volatile Compound Analysis). Table 1. GC–MS analysis of Prangos ferulacea (L.) L ndl. seed essential oil. defined components. 6 Chemical composition and gas of Prangos ferulacea (L.) L ndl. essential oil. The analyses were performed using chromatography-mass spectrometry (GC-MS). The analyses, AOC-20 automatic sampling system equipped with Sh madzu GCMS-QP 2010 Ultra It was implemented in the system. Automatic sampler settings, solvent prewashing, six The solvent included final washing and sample washing. Both absorption and injection 5 The p-stone velocities were set to high and the v-scosmic equalization time was 0.2 s. It was adjusted. The injection mode was set to normal. Gas chromatography separation, Rx-5MS capillary column (30.0 m length, 0.25 mm diameter, 0.25 µm film thickness) This was achieved using [method]. Gas flow control in pressure mode, total flow rate 10 The system was activated. The purging flow rate was set to 3.0 mL / min. Mass spectrometric detection is performed using electron ionization (EI) mode. It was realized. The ion source temperature was reached 200 °C and the interface temperature was reached 250 °C. It has been adjusted. Solver delay time recorded as 3 minutes. MS data, 0.30 s scan time and. 1428 scans were obtained in scanning mode with a speed of 40–450 m / z. Detector 15 The voltage was adjusted according to the automatic adjustment results of the device. Volatile components n identification, comparison of the obtained mass spectra with the reference mass spectra This was done by comparing the libraries. The results are in Table 1. It has been summarized. Creation of a CLP-derived sepsis model 20 Rats were distributed in groups of eight. The experimental study groups were: I. Sham group: The appendix was ruptured but not ligated or perforated. Group II CLP: Appendix Tie-and-Perforation Model III. CLP-PF 50 group: Appendix Tie-Off and Perforation Model +Prangos ferulacea (L.) L ndl. essential oil (PFO) 50 mg / kg 25 IV. CLP-PF 100 group: Appendix Tie-Off and Perforation Model +Prangos ferulacea (L.) L ndl. essential oil (PFO) 100 mg / kg V.CLP-PF 200 group: Appendix Tie-Off and Perforation Model +Prangos ferulacea (L.) L ndl. essential oil (PFO) 200 mg / kg VI. CLP-CEF 100 group: Cetruncatine Ligation and Perforation Model + Ceftr axon 30 100 mg / kg Sepsis model induced by cecal ligation and puncture (CLP) method (M shra et al., 2008). Rats were administered Xylaz n (10 mg / kg) and ketam n (90 mg / kg) under anesthesia. 7 The abdominal area is shaved and treated with R vanol, following asepsis and antisepsis rules. Accordingly, an incision 2 cm long was made on the left side of the medial line. Through the incision, the muscles will be retracted to reach the peritoneal membrane, and the peritoneum will be... The cecum was removed by opening it. The size of the cecum was measured in 3 / 5 ratio. From a distance of 3, using a numbered pl k, the l gature was ed. The apex of the cecum is “+” 5 The hole will be pierced in the direction indicated by the 20G needle tip. The contents will be drawn through the holes with a certain amount of pressure. After it comes out, it is placed back into the abdomen and pressure is applied to the peritoneum, muscles and skin in that order. The procedure was completed and the area was closed. The rats were placed in cages to keep them occupied while they came out of anesthesia. He will be taken to the area. In the sham group, an incision was made on the left side of his abdomen. The skin, muscles, and dermis were closed by applying pressure to the area. Rats, Prangos 10 Essential oil obtained from ferulacea (L.) L ndl. in doses of 100, 200, 400 mg / kg The saline solution was diluted and administered via a 1ml syringe starting from hour 0. It was administered intraperitoneally ( .p.) over 12 hours. The study was performed on rats at 36 hours. Anesthesia was administered by intraperitoneal ketamine, NH3 chloride, and KS2L NH3 chloride. Terminated. Each sample was subjected to cytopathological and immunohistochemical stock analysis at a rate of 10-15%. It was stored in formalin solution. Findings Prangos ferulacea (L.) L ndl. F tok myasal Prof l of Essential Oil The study used Gas Chromatography and Mass Spectrometry (GC-MS) methodology. performed using Prangos ferulacea (L.) L ndl. EO 'nf full myasal 20 The profile was evaluated. The main components of the essential oil include .gamma.- Terp nene (23.39%), cs-Oc mene (19.39%) and beta.-Oc mene (18.67%) It is included. Among the detected secondary components, trans-.beta.- accounts for 9.15%. Oc mene % 5.28 le Cymene <para->And .beta.-Phellandrene came out on top with 4.97%. SEM Findings 25 In the sham group, the alveolar surface was smooth and the Kohn's apertures were regular. However, A small number of erythrocytes were found on the alveolar surface. In the CLP group, alveolar erythrocytes were also observed. Pneumoderms were prominent on the surface, and separations were observed in places. CLP-PF50 In this group, the interpneumothorax was not prominent, and the Kohn's gaps were not. Pneumostats were prominent in the CLP-PF100 group. Kohn apertures were shown in sham 30. It resembled the group. Additionally, there were erythrocytes on the alveolar surface. CLP- In the PF200 group, the line of sight between the pneumostats had been lost, Kohn apertures. Although it appears mild, it is generally similar to the sham group. Inside the alveoli 8 Pneumatic strains were observed. Slight separations were observed among pneumatic strains in the CLP-CEF 100 group. (Figure 2). The CLP-PF50 and CLP-CEF100 groups showed statistically significant differences. was significantly lower (Figure 3A, P<0.001). The length of Kohn apertures, CLP-PF50 It was determined that the group decreased compared to the CLP, CLP-PF100 and CLP-CEF100 groups. (P<0.001). Also, the CLP-PF100 group was 5 times more active than the Sham and CLP-PF200 groups. It was noticed that it was long (Figure 3B. P<0.01). H. stomatological findings In the sham group, mild cell infiltration in the interalveolar septum, hyperemia, and Perbronchial-bronchial lymphoma tissue hyperplasia was observed. CLP group lung In the pathological examination of the tissue, hyperemia was found in the interalveolar septum, 10 Mononuclear and neutrophil cell infiltration in perbronchial-bronchial lymphoid tissue. There was hyperplasia and degeneration of the bronchi and bronchial epithelium. PF and CEF. In their groups, there was mild cell infiltration in the moderate to severe interalveolar septum, mild severe hyperplasia and mild hyperplasia in bronchial-bronchial lymphoid tissue (Figure 4A). Interalveolar 15 in CLP, CLP-PF100 and CLP-PF200 groups. Septum cell infiltration was determined to be more severe in the Sham group. (P<0.05). There was no statistically significant difference between the CLP-PF-50 and CLP-CEF100 groups. The results were slightly different (P>0.05). Hypertension was milder in the CLP-PF200 group compared to the CLP group. It was observed that (P<0.05). Perbronchial-bronchial lymphoid tissue hyperplasia, CLP and No statistically significant differences were found between the PF groups (Table 2, P>0.05). 20 Lung damage was increased in the CL group compared to the Sham group (P<0.001), CLP-PF (50- It was observed that the results were statistically similar in the 100-200 and CLP-CEF100 groups. (Figure 4B, P>0.05). Immunoh stock myasal Results In the interalveolar septum cell infection, in the perbronchus-bronchus lymphoid tissue, 25 NF-kB, TNF-α, IL-1β, IL-6 and HMGB1 in bronchial and bronchial cell walls The expression is summarized in Table 2. In interalveolar septum cell infiltration, HMGB1 expression was higher in the CLP-PF100 and CLP-PF200 groups compared to the CLP group. Expression was found to be decreased (P<0.05). In perbronchus-bronchus lymphatic tissue, Sham Compared to the group, CLP-CEF100 was found to be decreased (P<0.005). NF-kB 30 expression, in intraalveolar septum cell infiltration in the CLP-PF100 group, It was determined that CLP and CLP-CEF100 were decreased according to the groups (P<0.05). Perbronchial- NF-kB expression in bronchial lymphoid tissue was increased in the CLP-CEF100 group, CLP- 9 A decrease was detected in the PF50 and CLP-PF100 groups (P <0.05). Bronchi and bronchioles NF-kB expression in ep tel cells is higher in CLP-PF50 group and CLP-CEF100 group. TNF-α expression was found to be decreased compared to the group (P<0.05). Cellular influenza decreased in the CLP-PF50 group compared to the CLP group. bel rlend (P<0.05). Perbronchial-bronchiole lymphoid tissue and bronchial and bronchiole epithelium 5 TNF-α expression in cells was statistically significant between CLP and PF groups. No difference was found (Figure 15, P>0.05). Table 2. Histopathological and immunohistochemical changes in lung damage. stat st ksel olarak show m 15 25

Claims

REQUESTS 1. The invention relates to the prevention and / or treatment of sepsis-induced acute lung injury. It is an essential oil compound intended for use, and its properties are: Prangos ferulacea (l.) l ndl. b tk s , terc hen bu b tk nn essential oil çermesi le Karakter zed r. 5 2. Claim 1 states that the invention is gbbr and its characteristic is; Prangos ferulacea (l.) l ndl. volatile It is characterized by containing 100-400 mg / kg of oil.

3. Claim 1 states that the invention is gbbr and its characteristic is; Prangos ferulacea (l.) l ndl. volatile. The oil is characterized by containing preferably 100 mg / kg or 200 mg / kg.

4. Claim 1'k gbbr invention, its characteristic ğ; Prangos ferulacea (l.) l ndl. Volatile 10 The oil contains HMGB1, NF-κB, TNF-α, IL-1β and IL-6 pro-inflammatory mediators. Their expressions are characterized by their simultaneous occurrence of nh be. 20 30

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